New application of neodymium ion-sensitized upconversion nanocrystals and high-resolution multiphoton microscopy system

Abstract

The invention discloses a new application of neodymium ion-sensitized up-conversion nanocrystals and a high-resolution multiphoton microscope system. Neodymium ion-sensitized upconversion nanomaterials can be excited by short-wavelength steady-state lasers with a central wavelength of <800nm ​​to generate multiphoton visible light. They have larger multiphoton absorption cross-sections and multiphoton saturation excitation power, making it easier to perform high-order multiphoton imaging. This new feature can greatly reduce the cost of the system and significantly improve the resolution of microscopic imaging when used in multiphoton microscopic imaging. The multiphoton microscopic imaging system includes a short-wavelength steady-state laser with a central wavelength of <800nm, and its sample adopts an up-conversion nanometer material sensitized by neodymium ions. The present invention proposes for the first time the use of short-wavelength steady-state lasers to construct a cheap and simple multi-photon microscopy system, and proposes to use neodymium ions to sensitize nanomaterials with upconversion luminescent properties for ultra-high-resolution multi-photon microscopy imaging. Can be introduced into cells, tissues, or other substrates for high-resolution bioimaging.

Description

technical field [0001] The invention belongs to the technical field of optical microscopy, and in particular relates to a new application of neodymium ion-sensitized up-conversion nanocrystals and a high-resolution multiphoton microscopy system. Background technique [0002] In modern optical imaging techniques, probe-labeled fluorescence microscopy, especially laser scanning confocal microscopy based on fluorescence single-photon processes, has been achieved in the fields of medicine, life sciences, and materials science due to its high resolution (about 200 nanometers). Wide range of applications. However, confocal microscopy also has many shortcomings, such as relatively complex system, ultraviolet and blue light excitation is easy to damage biological samples, the imaging depth is very limited (tens of microns), it is difficult to avoid the interference of biological sample autofluorescence, and samples are easy to photobleach, etc. Based on the natural "optical section...

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Problems solved by technology

[0003] However, the nonlinear multiphoton (two-photon, three-photon, four-photon) excitation cross section of most fluorescent substances is very small, and the use of ultra-high peak power femtosecond pulsed lasers can obtain stronger multiphoton fluorescence, but the high cost of light sources ( ~$200,000) limits its widespread promotion, and the use of three-photon and four-photon fluorescence wavelengths under short-wavelength excitation usually falls in the ultraviolet band, which requires high detection optical paths and detectors, which is not conducive to short-wavelength excitation. Multiphoton High Resolution Microscopy

Although low-cost two-photon microscopy with steady-state laser pumping has been proposed, the resolution of low-order two-photon imaging excited by near-infrared wavelengths (greater than 800 nm) is still not high

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